(A and B) TRPV1 mRNA was detected by RT-PCR, and TRPV1 protein level was detected by immunoblotting in mesenteric arteries (MA) and cultured endothelial cells (ECs) from WT but not from
TRPV1−/− mice. M, marker. (C) Representative immunofluorescence images of TRPV1 in ECs (left panel, bar denotes 50 μm) and in mesenteric arteries (MA) (right panel, bar denotes 100 μm) from WT (upper panel) and
TRPV1−/− mice (lower panel). DAPI denotes visualizing nuclei. Coexpression of CD31 (red) with TRPV1 (green) is shown. (D) Representative tracings, with inset summary data showing capsaicin (CAP) stimulated increases of [Ca2+]i in ECs; **p < 0.01 versus capsaicin 1 nM, ##p < 0.01 versus capsaicin 0.1 μM. Data are means ± SEM (six separate experiments). (E) Representative tracings, with inset summary data showing the inhibitory effects of specific TRPV1 antagonist 5′-iodo-resiniferatoxin (iRTX), the intracellular Ca2+ chelator, BAPTA-AM and TRPV1 deficiency (
TRPV1−/−) on capsaicin-induced Ca2+ entry in cultured ECs. **p < 0.01 versus control. Data are means ± SEM (six experiments). (F) TRPV1 protein expression in mesenteric arteries (MA) from WT mice fed with normal diet (ND) and normal diet plus capsaicin (NC) for 6 months; *p
Figure 2. Activation of TRPV1 Increases Intracellular…
Figure 2. Activation of TRPV1 Increases Intracellular Calcium and Enhances eNOS Activity in the Vasculature
Figure 2. Activation of TRPV1 Increases Intracellular Calcium and Enhances eNOS Activity in the Vasculature (A) Colocalization of eNOS and TRPV1 in mesenteric arteries from WT (upper panel) and TRPV1-deficient mice (TRPV1−/−, lower panel), which were detected by immunofluorescence staining for eNOS (red) and TRPV1 (green). Scale bar denotes 100 μm. Photomicrographs are representative of three samples performed for each combination. (B and C) Immunoblottings showing the effect of capsaicin (1 μM) on the levels of p-eNOS and eNOS in cultured ECs from WT and from TRPV1−/− mice. *p < 0.05 versus control. Data are means ± SEM. Each n = 3. (D and E) Immunoblottings showing the effect of dietary capsaicin (0.01%) to WT and TRPV1−/− mice for 6 months on levels of p-eNOS and eNOS in mesenteric arteries (MA); *p < 0.05 versus ND. ND, normal diet; NC, normal diet plus capsaicin. The pooled data are means ± SEM. Each n = 3. (F) Levels of p-eNOS and eNOS by immunoblotting and summary data showing inhibition of capsaicin-induced increase of p-eNOS by specific TRPV1 antagonist 5′-iodo-RTX (iRTX), or the intracellular Ca2+ chelator BAPTA-AM, or EGTA in cultured ECs from WT mice; **p < 0.01 versus control, ##p < 0.01 versus capsaicin group. CAP, capsaicin. Data are means ± SEM. Each n = 3.
Figure 3. TRPV1-Mediated NO Production Is Associated…
Figure 3. TRPV1-Mediated NO Production Is Associated with Calcium Influx
(A and B) Representative pictures…
Figure 3. TRPV1-Mediated NO Production Is Associated with Calcium Influx (A and B) Representative pictures and summary data showing quantification of capsaicin-induced NO production in ECs using a dye 4,5-diaminofluorescein diacetate (DAF-2DA), which is converted into the fluorescent DAF-2T in the presence of NO. RTX, resiniferatoxin; iRTX, 5′-iodo-resiniferatoxin. *p #p < 0.05 versus capsaicin group. Scale bar denotes 100 μm. (C and D) Representative tracings and summary data showing the effects of iRTX, BAPTA-AM, and EGTA on capsaicin-induced NO production in cultured ECs. **p #p < 0.05 versus CAP 0.1 μM. (G and H) Representative tracings and summary data showing the effects of L-NAME on capsaicin-induced NO production in mesenteric arteries from WT mice; **p ##p < 0.01 versus TRPV1−/− mice; ΔΔp < 0.01 versus TRPV1 heterozygote mice (TRPV1+/−). Data are means ± SEM (three to six experiments).
Figure 4. Effects of TRPV1 Activation on…
Figure 4. Effects of TRPV1 Activation on PKA Phosphorylation and Activity
Figure 4. Effects of TRPV1 Activation on PKA Phosphorylation and Activity (A) The effect of TRPV1 activation on PKA activity in cultured ECs. CAP, capsaicin; EGTA, ethylene glycol tetraacetic acid; iRTX, 5′-iodo-RTX; *p TRPV1−/− mice. Data are means ± SEM. Each n = 3. (G) The effect of chronic TRPV1 activation on PKA phosphorylation in mesenteric arteries (MA) from WT mice. ND, normal diet; NC, capsaicin diet; *p TRPV1−/− mice. Data are means ± SEM (four to eight arterial rings).
Figure 5. Chronic Activation of TRPV1 Enhances…
Figure 5. Chronic Activation of TRPV1 Enhances Endothelium-Dependent Relaxation
(A) Effect of endothelium denudation on…
Figure 5. Chronic Activation of TRPV1 Enhances Endothelium-Dependent Relaxation (A) Effect of endothelium denudation on capsaicin-induced relaxation in freshly isolated mesenteric arteries from WT mice; *p TRPV1−/−) on capsaicin-induced relaxation in mesenteric arteries; *p < 0.05 versus WT mice. (C and D) Acetylcholine- and nitroglycerin-induced relaxation in the presence or absence of L-NAME (100 μM) in isolated mesenteric arteries from WT mice on normal diet (ND) or capsaicin diet (NC); *p TRPV1−/−) on normal diet (ND) or capsaicin diet (NC). Data are means ± SEM (four to eight arterial rings).
Figure 6. Role of Perivascular Sensory Nerves…
Figure 6. Role of Perivascular Sensory Nerves in TRPV1-Mediated Relaxation
(A) Representative tracings showing inhibitory…
Figure 6. Role of Perivascular Sensory Nerves in TRPV1-Mediated Relaxation (A) Representative tracings showing inhibitory effect of L-NAME and CGRP receptor antagonist CGRP 8-37 on capsaicin-induced relaxation in mouse mesenteric arteries. (B) Effect of L-NAME (100 μM) on capsaicin-induced relaxation in freshly isolated mesenteric arteries from C57BL/6J mice; **p
Figure 7. Effects of TRPV1 Activation on…
Figure 7. Effects of TRPV1 Activation on Blood Pressure and Vascular Relaxation in Spontaneous Hypertensive…
Figure 7. Effects of TRPV1 Activation on Blood Pressure and Vascular Relaxation in Spontaneous Hypertensive Rats (A) The plasma concentration of capsaicin in rats administered intragastrically with a single dose of capsaicin (15 mg/kg body weight). (B–D) Identification of capsaicin in plasma: mass spectrometry of capsaicin, chromatogram of extracted plasma sample from capsaicin-treated rats, and chromatogram of authentic capsaicin. (E) The time course of capsaicin-induced reduction in SBP in SHR fed with normal diet (ND) or normal diet plus capsaicin (NC) for 7 months. SBP was determined using tail cuff method. *p
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Publication types
Research Support, Non-U.S. Gov't
MeSH terms
Capsaicin / pharmacology*
Cyclic AMP-Dependent Protein Kinases / metabolism
Endothelial Cells / enzymology
Endothelial Cells / metabolism
Endothelium, Vascular / cytology
Endothelium, Vascular / physiology*
Hypertension / prevention & control*
Nitric Oxide / metabolism
Nitric Oxide Synthase Type III / metabolism
TRPV Cation Channels / deficiency
TRPV Cation Channels / genetics
TRPV Cation Channels / metabolism*
Substances
Nitric Oxide Synthase Type III
Cyclic AMP-Dependent Protein Kinases
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